A. Field of the Invention
This invention relates to specifically designing and genetically engineering recombinant baculovirus for producing, in a compatible insect system, a desired protein, virus, protein hybrid, or virus hybrid. In particular aspects, this invention relates to the use of different baculovirus promoters for the ultimate purpose of constructing a recombinant baculovirus designed for the investigator's specific need.
B. Description of the Related Art
The capabilities of producing genetically engineered proteins, through the use of recombinant DNA technologies, have dramatically increased in the last decade. However, problems have accompanied these new capabilities. One of the many problems involves constructing and genetically engineering the desired protein product. The operative words in the last sentence are "engineering the desired protein product." All too often, when it comes to creating a construct specifically designed for the production of a desired protein or virus, comprises have to be made. It is not uncommon that many intervening and tedious steps are involved in the creation of a desired construct for the purpose of producing a desired product. It is also not uncommon to have "to settle" for the production of an almost perfect construct because of technical difficulties in designing compatible sites for insertion of desired genes.
It is the intent of this invention to detail methods and compositions for designing and creating a baculovirus construct. This construct is designed in such a manner that few if any compromises have to be made in order to create the desired product. This baculovirus construct allows for the production of virtually any protein, vaccine, therapeutic, diagnostic, viral pesticide, or virus, as well as, the creation of virtually any new combination of the above. Combinations of proteins or viruses are termed "protein hybrid" or "virus hybrid." These terms imply there are multiple sources from which the product is created or that the product itself is made up of different components.
This invention will allow the investigator to design and create desired proteins, vaccines, viruses, therapeutics, diagnostics, viral pesticides, hybrids of the above, or functional and active domains from any of the above as well. Therefore, the potential use of this vector is endless.
The success of this invention, is in part, due to the unique use of multiple baculovirus promoters (both early and late) along with homologous recombination into a non-essential region(s) of the AcMNPV genome. For optimal flexibility, this invention employs three baculovirus promoters: the natural polyhedrin promoter mapped to the Eco-RI site of Fragment I; an early baculovirus promoter and a late baculovirus promoter.
The present invention also describes the production of recombinantly generated virus or virus hybrid particles or proteins involving the novel use of an early baculovirus gene promoter to temporally drive the expression of truncated or mutated non-structural genes of a virus particle as a unit, 5' to a late baculovirus gene promoter controlling the expression of cDNA for structural genes from the same or a different virus. The recombinantly expressed virus or virus hybrid, obtained from this construct, is essentially intact, virtually identical to the authentic parent virus particle, highly antigenic yet non-replicative and is therefore exceedingly functional in terms of use as a vaccine or diagnostic reagent.
This invention allows the investigator the flexibility to design and create a baculovirus construct that produces either a desired protein, vaccine, virus, therapeutic, diagnostic, viral pesticide, hybrid of the above, or functional and active domain from any of the above as well. This invention will be accomplished efficiently and without undue experimentation.